人类白细胞抗原DR分子和细胞间粘附分子—1在正常和病变角？…

目的 探讨正常及病变角膜中人类白细胞抗原ＲＤＲ分子和细胞间粘附分子－１的表达及其在角膜炎症和移植排斥反应中的作用和影响。方法 应用免疫组化技术对１０例正常信 患者的病变角膜标本进行人类白细胞抗原ＤＲ分子和细胞间粘附分子－１的表达的检测。结果正常人角膜无或轻微表达人类白细胞抗原ＤＲ分子和细胞粘附分子－１；病变角膜，尤其是炎症角膜及角膜移植术后发生排斥反应的角膜，基创性表达率明显增高。结论 人类白细胞     

FK506抑制大鼠角膜移植免疫排斥反应的免疫病理学研究

目的 研究角膜移植免疫排斥反应的免疫病理学变化及免疫细胞和某些相关免疫分子在角膜移植免疫排斥反应中的作用,阐明了ＦＫ５０６的免疫抑制机理。方法 应用免疫组织化学染色方法检测ＦＫ５０６,环胞霉素Ａ及对照组角膜植片中ＣＤ＾＋４,ＣＤ＾＋８,巨噬细胞,白细胞介素－２受体,Ⅱ类主要组织相容性抗原,细胞间粘附分子－１及淋巴细胞功能相关抗原－１的表达。结果 排斥的角膜组织大表达上述免疫细胞和分子,ＦＫ５０６可     

γ—干扰素诱导角膜上皮细胞和内皮细胞HLA抗原在体外表达的实验研究

目的了解γ－干扰素对角膜细胞ＨＬＡ抗原表达的影响。动态观察角膜细胞间ＨＬＡ－ＡＢＣ抗原和ＨＬＡ－ＤＲ抗原表达的量变关系。方法采用ＡＣＡＳ－５７０粘附式细胞分析仪和间接免疫荧光技术，对体外原代培养的正常成人角膜上皮和内皮细胞ＨＬＡ－ＡＢＣ位点和ＨＬＡ－ＤＲ位点抗原表达的相对量进行测定。结果在体外，γ－干扰素可诱导角膜上皮细胞和内皮细胞ＨＬＡ－ＤＲ抗原的异常表达，同时使ＨＬＡ－ＡＢＣ位点抗原表达量增加；经γ－干扰素诱导后，上皮细胞的ＨＬＡ－ＡＢＣ抗原及ＨＬＡ－ＤＲ抗原的表达量均较相应的内皮细胞表达量高；且两种细胞的ＨＬＡ－ＡＢＣ抗原表达量增高幅度较小，而ＨＬＡ－ＤＲ抗原增高幅度较大。结论角膜细胞ＨＬＡ抗原的异常表达可能是角膜移植排斥反应发生的主要原因；γ－干扰素在角膜移植排斥反应发生时充当重要角色。     

细胞间粘附分子在角膜疾病中的表达

目的：探讨细胞间粘附分子（Intercellular Adhesion Molecule-1,ICAM-1）在感染性角膜疾病和角膜移植免疫排斥中的表达及作用。方法：应用抗细胞间粘附分子ICAM-1的单克隆抗体，对42例角膜移植所取下的病变角膜进行免疫组织化学研究。角膜组织连续冰冻切片，进行SABC免疫组化及HE染色，研究细胞间粘附分子的表达与炎性细胞分布以及炎症程度的关系。结果：各种感染性角膜疾病、角膜变性以及角膜移植排斥过程中，ICAM-1在角膜上皮，尤其是基底细胞、角膜细胞、内皮细胞和血管化的基质层新生血管内皮细胞中均有表达，在炎性细胞包括单核细胞、巨噬细胞及淋巴细胞浸润部位表达增强；ICAM-1表达强度与炎症程度相平行。结论：ICAM-1在感染性、变性角膜病及角膜移植排斥中均有表达，在炎性细胞浸润部位表达增强；I-CAM-1对白细胞募集于炎症部位、炎症的发生、发展具有重要的中介作用。     

蚕蚀性角膜溃疡角膜及邻近球结膜的免疫组织化学研究

用免疫组化技术对１４例蚕蚀性角膜溃疡的病变角膜及邻近球结膜标本的人类白细胞ＤＲ抗原（ＨＬＡ－ＤＲ）表达及Ｔ淋巴细胞亚群进行检测，发现有大量的角膜和球结膜的上皮细胞、角膜基质细胞异常地表达ＨＬＡ－ＤＲ抗原，同时，辅助性Ｔ细胞／抑制性Ｔ细胞（Ｔ＿Ｈ／Ｔ＿Ｓ）比例较正常对照组明显增高。结果提示，周边部角膜和球结膜主要组织相容性Ⅱ类抗原（ＭＨＣ－Ⅱ）的异常表达以及Ｔ＿Ｈ／Ｔ＿Ｓ升高导致过强的自身免疫反应，可能是本病发病的直接原因。     

Leflunomide抑制大鼠角膜移植排斥反应的免疫病理学研究

目的：研究角膜移植排斥反应的免疫病理学变化，阐明Ｌｅｆｌｕｎｏｍｉｄｅ的免疫抑制机理。方法：应用免疫组织化学染色方法检测环孢霉素Ａ、Ｌｅｆｌｕｎｏｍｉｄｅ及对照组角膜移植片中ＣＤ４＾＋细胞、ＣＤ８＾＋细胞、白细胞介素－２受体及主要组织相容性Ⅱ类抗原的表达。结果：排斥的角膜组织大量表达上述免疫细胞和分子，Ｌｅｆｌｕｎｏｍｉｄｅ可以对这些细胞和分子产生明显的抑制作用。结论：迟发型超敏反应和细胞毒性Ｔ细     

抗ICAM-1单克隆抗体抑制大鼠角膜移植排斥反应的实验研究

目的 :探讨应用抗细胞间粘附分子 (Intercellularadhesionmolecule 1,ICAM 1)单克隆抗体中和治疗大鼠角膜移植排斥反应的效果。方法 :应用两种主要组织相容抗原完全不同的大鼠Louis和RCS鼠 ,建立近交系大鼠穿透性角膜移植动物模型 ,分三组给药 ,其分别是抗细胞间粘附分子单克隆抗体组 (抗ICAM 1抗体组 )、生理盐水组和环胞霉素A(CsA)组。观察角膜移植排斥发生的时间及治疗效果。标本作HE染色及免疫组化染色 ,观察免疫排斥反应的程度及炎症程度。结果 :三组角膜植片平均存活时间为 :生理盐水组角膜移植排斥反应平均发生时间为术后 12 .1天 ,CsA组为18.3天 ,抗ICAM 1抗体组为 2 0 .2天。HE染色显示抗ICAM 1抗体组及CsA组的炎症反应较生理盐水组明显减轻 ,免疫组化染色显示抗ICAM 1抗体组和CsA组的ICAM 1表达较生理盐水组明显减少。结论 :抗ICAM 1单克隆抗体中和治疗可减少移植角膜ICAM 1表达及免疫炎性细胞的浸润 ,减轻角膜移植免疫排斥反应 ,延长移植片的存活时间 ,是一种有效的新型免疫抑制剂     

细胞间黏附分子与角膜移植免疫

角膜移植免疫是多种免疫细胞和免疫分子参与的复杂的免疫反应过程。近来研究表明ICAM1在角膜移植免疫排斥反应的发生机制中起重要作用。ICAM1协同抗原递呈,促使免疫细胞在植片聚集,触发和促进移植排斥反应的发生和发展,引发粒细胞聚集于角膜基质层,导致组织损伤、破坏、角膜新生血管化使移植失败。目前研究抗ICAM1药物抑制角膜移植排斥反应已进入实验阶段。就细胞间黏附分子与角膜移植免疫的研究现状进行综述。     

Graves眼病患者眼外肌组织免疫组化研究

目的探讨Ｇｒａｖｅｓ眼病发生机制。方法应用单克隆抗体人类白细胞ＤＲ抗原（ＨＬＡＤＲ）、巨噬细胞（ＣＤ６８）、第八因子相关性抗原（Ｆ８／８６），ＣＤ３，ＣＤ４，ＣＤ８，Ｂ细胞（ＣＤ２０）对１４位Ｇｒａｖｅｓ眼病患者眼外肌标本进行免疫组化研究。结果患者眼外肌间质中有大量的ＤＲ＋细胞，有大量巨噬细胞浸润，血管丰富、扩张，淋巴细胞浸润，以ＣＤ３＋细胞为主。结论提示患者眼外肌间质中ＨＬＡＤＲ抗原的异常表达在该病的发生发展中起重要作用。细胞免疫是局部炎症的重要原因。     

角膜移植免疫排斥反应的免疫病理学研究

目的通过分析角膜移植排斥反应的免疫病理改变，探讨移植失败的相关因素。方法角膜移植术后排斥反应的患者42例，于再次行穿通性角膜移植术时取其排斥的移植片，进行免疫组化及HE染色，观察术后排斥反应的移植片中巨噬细胞、CD4^＋和CD8^＋细胞的浸润和分布、MHC-Ⅱ类主要组织相容性抗原、白介素受体-2和转化生长因子-β的表达，以及移植片的组织病理学改变。结果42例移植片均有上述免疫细胞的表达，并以新生血管周围更加明显；这些细胞表达MHC-Ⅱ类抗原和IL-2R；移植片的上皮和基质细胞中也有MHC-Ⅱ类抗原的表达；角膜上皮细胞可表达TGF-D；所有移植片内皮细胞均不完整甚至大部缺如。结论新生血管使免疫细胞进入移植片，破坏免疫赦免机制，与移植片免疫损害密切相关；免疫细胞和分子直接导致移植片免疫损伤，内皮细胞的严重损伤终将导致移植失败。     

HLA antigenicity of normal and pathological corneas

Fresh human corneas and corneal buttons were studied for expression of HLA antigens. Using monoclonal antibodies in an indirect immunofluorescence assay, corneal layers were examined for class I (HLA-A, B, C) and class II (HLA-DR) histocompatibility antigens. Twenty-one human corneas were studied, 6 normal and 15 pathological: 4 buttons of allograft rejection, 9 buttons of pseudophakic bullous keratopathy. In fresh control corneas, HLA-A, B, C antigens were localized on corneal epithelium and on stromal keratocytes but were never found on endothelial cells. HLA-DR antigens were not detected on corneal epithelium, stroma or endothelium but were detected on Langerhans cells within epithelium and anterior stroma. At the corneal limbus, HLA class I-II antigens were expressed on vascular endothelium. HLA antigen distribution was modified in pathological corneas. Antigens HLA-A, B, C were induced on endothelial cells of rejected corneal allografts. Antigens HLA-DR were detected on epithelial cells, cells in the stroma of pseudophakic bullous keratopathy and also on endothelial cells of rejected corneal allografts. These results suggest that induction of class I and II antigen expression by inflammatory factors may occur in vivo. In rejected corneal allografts induction of HLA-DR antigen on corneal layers would intensify the process of rejection. This study and others have demonstrated the ability of modulation of HLA antigen expression on human corneal cells in vivo.     

In situ immunohistochemical analysis of cell adhesion molecules on human corneal endothelial cells.

Interaction of leucocytes with human corneal endothelial cells (HCECs) can be observed in several clinicopathological conditions, such as uveitis, keratitis, and corneal graft rejection. Since leucocyte-endothelial cell interactions involve various adhesion receptors we have analysed the expression and distribution pattern of the neural cell adhesion molecule (NCAM), the intercellular adhesion molecule-1 (ICAM-1), the vascular cell adhesion molecule-1 (VCAM-1), the endothelial leucocyte adhesion molecule-1 (ELAM-1), and the cluster of differentiation antigen-44 (CD44) on flat preparations of normal and organ-cultured HCECs. NCAM and ICAM were constitutively expressed on HCECs whereas VCAM-1, ELAM-1, and CD44 were absent from normal HCECs. However flat mounts of HCECs from organ-culture preserved corneas showed a mosaic-like distribution pattern of VCAM-1 and ELAM-1 positive cells and garland-like clusters of CD44 positive cells. We suggest that modulation of ELAM-1, VCAM-1, and CD44 expression on HCECs may contribute to the regulation of leucocytes-HCECs interaction in the case of anterior segment inflammation.     

Proteinase and growth factor alterations revealed by gene microarray analysis of human diabetic corneas

PURPOSE: To identify proteinases and growth factors abnormally expressed in human corneas of donors with diabetic retinopathy (DR), additional to previously described matrix metalloproteinase (MMP)-10 and -3 and insulin-like growth factor (IGF)-I. METHODS: RNA was isolated from 35 normal, diabetic, and DR autopsy human corneas ex vivo or after organ culture. Amplified cRNA was analyzed using 22,000-gene microarrays (Agilent Technologies, Palo Alto, CA). Gene expression in each diabetic corneal cRNA was assessed against pooled cRNA from 7 to 9 normal corneas. Select differentially expressed genes were validated by quantitative real-time RT-PCR (QPCR) and immunohistochemistry. Organ cultures were treated with a cathepsin inhibitor, cystatin C, or MMP-10. RESULTS: More than 100 genes were upregulated and 2200 were downregulated in DR corneas. Expression of cathepsin F and hepatocyte growth factor (HGF) genes was increased in ex vivo and organ-cultured DR corneas compared with normal corneas. HGF receptor c-met, fibroblast growth factor (FGF)-3, its receptor FGFR3, tissue inhibitor of metalloproteinase (TIMP)-4, laminin alpha4 chain, and thymosin beta(4) genes were downregulated. The data were corroborated by QPCR and immunohistochemistry analyses; main changes of these components occurred in corneal epithelium. In organ-cultured DR corneas, cystatin C increased laminin-10 and integrin alpha(3)beta(1), whereas in normal corneas MMP-10 decreased laminin-10 and integrin alpha(3)beta(1) expression. CONCLUSIONS: Elevated cathepsin F and the ability of its inhibitor to produce a more normal phenotype in diabetic corneas suggest increased proteolysis in these corneas. Proteinase changes may result from abnormalities of growth factors, such as HGF and FGF-3, in DR corneas. Specific modulation of proteinases and growth factors could reduce diabetic corneal epitheliopathy.     

Immunohistochemical Expression of Fas Ligand (FasL) and Neprilysin (Neutral endopeptidase/CD10) in Keratoconus

Recent evidence demonstrated a correlation between apoptosis and neprilysin expression. The aim of this study was to investigate the immunohistochemical expression of Fas ligand (FasL) and neprilysin in keratoconic corneas in comparison to normal cadaver corneas to evaluate if such molecules play a role in the pathogenesis of keratoconus. We studied the expression of FasL and neprilysin in corneal specimens removed during penetrating keratoplasty in 15 cases with keratoconus and compared them with 5 normal cadaver corneas. In keratoconus, FasL was expressed in epithelium, endothelium and sub-Bowman’s stroma only, while neprilysin was expressed in epithelium, endothelium and all stromal layers. All normal corneas showed weak expression of both markers in basal epithelial layer only. In keratoconus, corneal epithelium with higher expression of FasL may evoke apoptosis in keratocytes, while neprilysin could prevent possible rescue of keratocytes from apoptosis.     

Detection of HLA class I and II antigens in rejected human corneal allografts

We compared the distribution of HLA-ABC (class I) and HLA-DR (class II) antigens on fresh human donor corneal tissue, donor corneas following a 72-hour storage in McCarey-Kaufman (M-K) medium, and corneal buttons from patients with allograft rejection and with chronic herpetic stromal keratitis. Incubation in M-K media had little or no effect on the distribution of HLA antigens as compared with fresh tissue. In contrast to control corneas, both HLA class I and II antigens were detected on corneal endothelial cells, cells in the stroma, and on basal epithelial cells in rejected allografts. Corneal endothelium in herpetic buttons did not express detectable HLA antigens. HLA-DR positive Langerhan's cells were demonstrated in the central corneal epithelium of rejected allografts, as well as in herpetic corneas, but not in control corneas except at the limbus. Based upon these observations, a theory of corneal allograft rejection in humans is proposed based upon the induction of class I HLA-ABC and class II HLA-DR antigens on cells in the donor button by a factor(s) associated with cellular inflammation.     

Expression of adhesion molecule CD44 on human corneas

AIMS—This study was undertaken to confirm the distribution and expression of the molecule CD44 on human corneas under normal and pathological conditions.
METHODS—Fifty eight corneal buttons from adult patients suffering from various corneal diseases and four normal corneas were included in this study. Frozen sections were stained immunohistochemically with monoclonal antibodies against human CD44 using an APAAP method and observed under a light microscope.
RESULTS—In normal corneas CD44 was predominantly expressed on the membranes of basal epithelial cells and on the keratocytes, as well as on the vascular endothelial cells of the corneal limbi, but was not expressed on corneal endothelial cells. Enhanced expression of CD44 was observed on the epithelium of corneas with inflammation and allograft rejection. In a number of abnormal conditions including allograft rejection, corneal trauma, primary and secondary corneal endothelial decompensation the remaining endothelial cells stained positively for CD44. However, in some corneas of keratitis, keratoconus, and dystrophy the endothelium which appeared relatively integral in morphology and amount remained CD44 negative.
CONCLUSIONS—These results suggest that CD44, the hyaluronate receptor, may play an important role in corneal cell-cell and cell-matrix interactions. Its regulation is closely related to corneal inflammatory reactions. The induction of CD44 on corneal endothelium might play a potential role in compensatory processes when corneal endothelial cells are injured.

     

Therapeutic keratoplasty using preserved corneas from keratoconus eyes

PURPOSE: To report and discuss cases of lamellar keratoplasty using corneas obtained during previous penetrating keratoplasty in keratoconus eyes. METHODS: Corneal buttons were obtained from 7 keratoconus patients and stored in a preserving solution for 7-60 days (average, 32.4 days) before use. The recipient eyes comprised recurrent pterygium 3 eyes, primary pterygium 1 eye, pseudopterygium 1 eye, corneal perforation with iris prolapse due to fungal corneal ulcer 1 eye, and limbal dermoid 1 eye. RESULTS: The recipient eyes ran favorable courses in general. Graft rejection developed in 2 eyes and was successfully treated with topical and systemic corticosteroid. CONCLUSIONS: Preserved corneas from keratoconus eyes were found useful in therapeutic lamellar keratoplasty. By this procedure, the current inadequate supply of donor corneas in eye banks in Japan can be augmented.     

干燥保存角膜在感染性角膜溃疡治疗中的应用

目的:评价应用干燥保存角膜片对554例严重感染性角膜溃疡行治疗性角膜移植手术的效果。方法:选自我院角膜病组1976-08/2000-12期间应用干燥保存角膜片对严重感染性角膜溃疡行治疗性角膜移植手术共计554例,本组病例统计分类为真菌性(238例)、单疱病毒性(170例)、细菌性(70例)及混合感染性(76例角膜溃疡4种,分析临床应用效果。结果:本组手术治疗严重感染性角膜溃疡平均有效率88.8%,其中真菌性角膜溃疡81.5%,单疱病毒性角膜溃疡98.2%,细菌性角膜溃疡97.1%,混合感染性角膜溃疡82.9%。结论:无水氯化钙-硅胶干燥长期保存的角膜片为临床随时提供角膜材料,是一项简单易行、便于推广应用的方法;在临床治疗过程中尽快、尽早查明感染病原体,可避免盲目性;对严重感染性角膜溃疡,治疗性角膜移植可直接清除病灶、较快地控制炎症、缩短治疗过程,达到保存眼球及恢复部分有用视力,为下一步光学性角膜移植创造条件。